45 3 Vol.45, No.3 2014 5 OCEANOLOGIA ET LIMNOLOGIA SINICA May, 2014 (Portunus trituberculatus) * ( 315211) (Portunus trituberculatus),, 25mg/kg,, 3P97 :, ;,, ;,,,,,,, 10d ; (Portunus trituberculatus); ; ; S948 doi: 10.11693/hyhz20130109001 (florfenicol),,, (Samuelsen et al, 1998; Stefan et al, 2004;, 2005;, 2010;, 2011;, 2012), (Oncorhynchus mykiss) (Pseudosciaena crocea) (Gadus morhua) (Salmo salar) (Acipenser baeri) (Anguilla anguilla) (Silurus asotus) (Crucian carp) (Fenneropenaeus chinensis) (Procambarus clarkii) (Penaeus vannamei Boone)(Pelodiscus sinensis) (, 2004; Park et al, 2006;, 2010;, 2011;, 2012), (Portunus trituberculatus),,,, (, 2002;, 2006),,,, (, 2010),, *, Y307391 ;, 2010R50025-10 ;, 2011B 81003 ;, 2011C50008 ;, XKL11093,, E-mail: zhaoqingsong@ nbu.edu.cn : 2013-01-09, : 2013-04-25
3 : 569 1 1.1 (P. trituberculatus), (90±25)g, 220,,, 25±1, (21±2) C, ph 8.0±0.5, 15d,, 50%, 1.2 ( 8g/L, 20g/L, EDTA 3.36g/L, 80 100μL/L); ( 10%, ); ( 99.2%, ) 1.3 Agilent Technologies1200 ( Agilent ), G1311A G1314B G1311A Chemstation G1329A ; R206D 1.4 (Vibrio alginolyticus) (2010) 1.5 (, 2011) 25mg/kg, 0.25 0.5 1 2 4 6 8 12 24 48 72 96 120 168 240h 1.. 1, 4 C 10min(3000r/min), ;, 20 C 5, 1.6, 0.5mL 0.5g, 4mL, 5min, 4 C 10min(6000r/min), 20mL,, 45 C, 1mL, 2min, 0.45μm, 20 C, (HPLC), 1.7 : Diaimonsil TM C18 (150mm 16mm i.d., 5μm); :.. = 27:73(v/v), 0.45μm ; : 1.0mL/min; : 223nm; : 30 C; ph 7.0; : 20μL 1.8 100μg/mL, 20 C 10 5 2 1 0.5 0.05μg/mL HPLC,,, S/N=2 (LOD) 1.9 0.5 1 5μg/mL(μg/g), 1.6, 3 (%)= / 100%3 31d3, 3d, 1.10 Microsoft Excel 2003 ; 3P97 2 2.1 0.05 10μg/mL (X)(Y), Y 32.51X+0.79, R 2 1 S/N=2, 0.025μg/mL 2.2 (84.8±1.6)% (88.0±8.9)% (94.2±2.0)%, 80%; 2.63 2.18 2.54, 3.67 3.23 4.61, 5%,, 2.3-1 3,,
570 45, 12h, 3.23μg/g, 1.64μg/g,, 2h, Fig.1 1 Change of florfenicol concentration with time in hepatopancreas of P. trituberculatus 2h 4h, 24h,, 168h 240h 0.1μg/mL, 98.6% 2.4 Fig.2 Fig.3 2 Change of florfenicol concentration with time in muscle of P. trituberculatus 3 Change of florfenicol concentration with time in plasma of P. trituberculatus 3P97, 1 2, (T 1/2β ) (CLs) (V d ), (T 1/2Ka ) (T peak ) (C max ) CLs V d, Ka T 1/2Ka 0.25h,,, 6.85μg/g 3.32μg/g;, 2h 15.16% 33.21%; T 1/2β T peak V d (P<0.05); Ka T 1/2Ka T peak C max AUC, Ka T 1/2β AUC, AUC CLs (P<0.01) Tab.1 表 1 氟苯尼考在健康和患病三疣梭子蟹组织内的药动学方程 Equations of florfenicol concentration with time in healthy and diseased P. trituberculatus R 2 R 2 C=9.22e 3.07t +1.48e 0.02t 10.70e 15.89t 0.97 C=19.28e 3.29t +1.42e 0.01t 20.71e 4.36t 0.88 C=6.75e 0.67t +1.44e 0.01t 8.19e 2.42t 0.97 C=19.01e 0.68t +1.44e 0.01t 20.45e 0.87t 0.97 C=15.23e 0.25t +0.69e 0.02t 15.92e 0.70t 0.99 C=18.07e 0.25t +1.71e 0.01t 19.78e 0.39t 0.99
3 : 571 表 2 氟苯尼考在健康和患病三疣梭子蟹体内的药动学参数 Tab.2 Pharmacokinetic parameters of florfenicol in healthy and diseased P. trituberculatus A(μg/mL) 9.222 a 6.753 b 15.230 19.283 d 19.012 e 18.071 B(μg/mL) 1.484 1.445 0.696 1.427 1.440 1.711 α(/h) 3.068 0.670 0.253 3.286 0.676 0.249 β(/h) 0.017 0.014 0.015 0.014 0.011 0.012 Ka(/h) 15.886 a 2.419 b 0.697 c 4.363 d 0.874 e 0.393 f K 10 (/h) 0.102 0.057 0.123 0.061 0.042 0.052 K 12 (/h) 2.460 0.464 0.114 2.472 0.469 0.150 K 21 (/h) 0.524 0.163 0.030 0.767 0.176 0.060 T 1/2Ka (h) 0.044 a 0.287 b 0.995 c 0.159 d 0.793 e 1.762 f T 1/2α (h) 0.226 1.034 2.741 0.211 1.025 2.783 T 1/2β (h) 39.903 a 50.236 b 46.609 c 48.437 d 63.349 e 55.667 f T peak (h) 0.143 a 0.816 b 2.385 c 0.331 d 1.662 e 3.700 f C max (μg/ml) 6.324 a 4.199 b 5.980 c 3.032 d 2.809 e 4.209 f AUC(μg/(mL h)) 87.765 a 111.396 b 84.153 c 100.843 d 136.329 e 159.649 f CLs(L/(kg h)) 0.285 a 0.224 b 0.297 c 0.248 d 0.183 e 0.157 f V d (L/kg) 2.802 3.957 b 2.408 c 4.043 d 4.366 e 3.017 f A: ; B: ; α: ; β: ; Ka: ; K 10 : ; K 12 : ; K 21 : ; T 1/2Ka : ; T 1/2α : ; T 1/2β : ; T peak : ; C max : ; AUC: ; CLs: ; V d : (P<0.05) 3,, ; 0.25h,,, (, 2001;, 2002;, 2012) (Park et al, 2006;, 2010;, 2011;, 2012;, 2012), (2011)(2006) (2007),,,,,, (, 2011;, 2006),, (Baggot, 1980) Uno(1996) ; (2006)(2011),, ;, (, 2003)(, 2001;, 2003)(, 1996),,,,,, (2006) (2003),,,,
572 45,,,,, V d,,, (, 2003) 0.1mg/kg, 1 3, 168h 240h, 0.1mg/kg,, 10d,,, 2010.., 25(4): 285 288,,, 2006.., 24(4): 526 231,,, 2012.., 21(40): 568 574,,, 2012.., 33(2): 235 238,,, 2006.., 30(4): 509 514,,, 2003.., 36(9): 1100 1104,, 2003.. :, 95 97,,, 2004.., 28(Supp1.): 63 68,,, 2012.., (2): 233 235,,, 2010.., 3(5): 116 121,,, 2011.., 30(2): 257 262, 2001. -. :,,, 2010.. ( ), 23(4): 35 40, 2002.., (3): 24 25,,, 2011.. ( ), 16(2): 92 96,,, 2007.. ( ), 20(1): 23 26,,, 2002.., 33(4): 384 388,,, 2005.., 12(4): 512 518,,, 2006.., 15(4): 448 455,,, 2003.., 34(1): 98 102,,, 2011.., (5): 42 43,,, 2012. EGCG (Pelteobagrus vachelli)., 43(5): 938 942,,, 2001.., 21(1): 86 89,, 1996.., 30(6): 10 15 Baggot J D, 1980. Distribution of antimicrobial agents in normal and diseased animals. Journal of the American Veterinary Medical Association (USA), 176(10): 1085 1090 Park B K, Lim J H, Kim M S et al, 2006. Pharmacokinetics of florfenicol and its metabolite, florfenicol amine, in the Korean catfish (Silurus asotus). Journal of Veterinary Pharmacology and Therapeutics, 29(1): 37 40 Samuelsen O B, Hjeltnes B, Glette J, 1998. Efficacy of orally administered florfenicol in the treatment of furunculosis in Atlantic salmon. Journal Aquat Anim Health, 10(1): 56 61 Stefan S, Corinna K, Axel C et al, 2004. Molecular basis of bacterial resistance to chloramphenicol and florfenicol. FEMS Microbiology Reviews, 28(5): 519 542 Uno K, 1996. Pharmacokinetic study of oxytetracycline in healthy and vibriosis-infected ayu (Plecoglossus altivelis). Aquaculture, 143(1): 33 42
3 : 573 PHARMACOKINETICS OF FLORFENICOL IN DISEASE MODEL OF PORTUNUS TRITUBERCULATUS ZHAO Qing-Song, JIN Shan, CHEN Yin-Er, QIN Fang-Jin, WANG Xi-Bo, WANG Guo-Liang (School of Marine sciences of Ningbo University, Ningbo 315211, China) Abstract To understand the metabolic differences of florfenicol in healthy and diseased Portunus trituberculatus, the pharmacokinetics of florfenicol were investigated based on Vibrio alginolyticus disease model. P. trituberculatus was orally administered in a single dose of 25mg/kg in water temperature (21±2) C. Tissues of haemolymph, muscle, and hepatopancreas were sampled and measured using high performance liquid chromatography (HPLC). Results show that florfenicol in healthy P. trituberculatus have a similar concentration-time curve with that of diseased animals. The profiles of the three tissues could be described by a two-compartment open model with a time lag. Florfenicol can be distributed unevenly in all tissues, absorbed most quickly in hepatopancreas and most slowly in plasma. Compared with the healthy P. trituberculatus, the rate of absorption and elimination in diseased P. trituberculatus slowed, the peak time delayed and half-liveextended, clean-up ratio and maximum drug concentration decreased, apparent volume of distribution and the area under the curve increased. These pharmacokinetic parameters showed that florfenicol in diseased P. trituberculatus changed greatly in absorption, distribution, elimination, and metabolism. According to the maximum residue limit (MRL) of 0.1mg/kg in tissues, the withdrawal period should not be less than 10 days under experiment condition. Key words florfenicol; Portunus trituberculatus; disease model; Vibrio alginolyticus; pharmacokinetics